Hematoxyllin-Eosin ( HE ) Staining

Freda Halim

Published: 2023-03-20 DOI: 10.17504/protocols.io.4r3l27zo3g1y/v1

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Abstract

This is a protocol to stain tissue with Hematoxyllin-Eosin ( HE ) Staining. We use human breast cancer tissue for running this protocol.

Steps

Deparaffinization

1.

Incubate slides in Xylol

2.

Incubate slides in Xylol

Rehydration

3.

Incubate slides in 100% Alcohol

4.

Incubate slides in 96% Alcohol

5.

Incubate slides in 80% Alcohol

6.

Incubate Slides in 70% Alcohol

7.

Rinse slides with Aquadest for 5 minutes

Nuclear Staining

8.

Incubate Slides in Hematoxyllin Reagent ( Hematoxyllin JT Baker )

9.

Rinse slides with running tap water

Defferentiation

10.

Incubate slides in bluing reagent @ 1 minutes, 5 times

11.

Rinse slides with running tap water

Cytoplasm staining

12.

Incubate slides in Eosin reagent 1 minute

Dehydration

13.

Incubate Slides in 70% Alcohol

14.

Incubate Slides in 80% Alcohol

15.

Incubate Slides in 96% Alcohol

16.

Incubate Slides in 100% Alcohol

Clearing

17.

Incubate Slides in Xylol

18.

Incubate slides in Xylol

19.

Incubate Slides in Xylol

Mounting and Finishing

20.

Wipe excess fluid,cover the slides with cover glass

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